中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2026, Vol. 21 ›› Issue (7): 2884-2892.doi: 10.4103/NRR.NRR-D-24-00435

• 原著:脑损伤修复保护与再生 • 上一篇    

化学和慢病毒方法生成人类诱导多能干细胞运动神经元的比较

  

  • 出版日期:2026-07-15 发布日期:2025-10-21

Comparative analysis of chemical and lentiviral approaches in the generation of human induced pluripotent stem cell–derived motor neurons

Masood Sepehrimanesh1, †, Wu Xu2 , Baojin Ding1, *   

  1. 1 Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center at Shreveport, Shreveport, LA, USA;  2 Department of Chemistry, University of Louisiana at Lafayette, Lafayette, LA, USA  †Current address: School of Biological Sciences, Louisiana Tech University, Ruston, LA, USA
  • Online:2026-07-15 Published:2025-10-21
  • Contact: Baojin Ding, PhD, Baojin.ding@lsuhs.edu.
  • Supported by:
    This work was supported by the National Institute of Health (NIH) National Institute of Neurological Disorders and Stroke (NINDS), Nos. NS112910, NS133252 (to BD); Department of Defense (DoD) Peer Reviewed Medical Research Program (PRMRP) Discovery Award, No. W81XWH2010186 (to BD).

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摘要：

人类诱导多能干细胞来源运动神经元（MNs）的生成克服了人类组织获取途径的限制，为肌张力障碍和肌萎缩性脊髓侧索硬化等运动神经元疾病的建模提供了前所未有的方法。通过不同策略生成的运动神经元在纯度、成熟度、表征甚至神经元特性方面都可能表现出很大差异，从而导致疾病建模和药物筛选的结果各不相同。然而，很少有人进行比较研究来确定通过不同方案制备的运动神经元之间的异同。实验通过慢病毒递送转录因子（Lenti-MNs）和化学诱导（ciMNs）方法制备了人类诱导多能干细胞来源运动神经元，并进行了系统的比较分析。结果发现，这两种方法生成的运动神经元都表现出典型的运动神经元形态，并能强有力地表达运动神经元特异性标记，如核同源转录因子 9和胆碱乙酰转移酶。ciMNs 方案利用小分子组合诱导运动神经元分化，成熟时间明显缩短，仅为 35 d，而 Lenti-MNs 为 46 d。此外，Lenti-MNs 适用于下游生化分析，而 ciMNs 则更适用于治疗方法，因为它们避免了慢病毒的使用。这两种方法都能生产出高纯度（>95%）和高产量的运动神经元。ciMNs 和 Lenti-MNs 在运动神经元标记和成熟标记方面没有发现明显差异。这些可靠的方法为研究人员提供了研究运动神经元疾病和潜在治疗策略的有力工具。

https://orcid.org/0000-0002-2149-2599 (Baojin Ding)

关键词: 化学物质, 人类诱导多能干细胞, 慢病毒, 运动神经元疾病, 运动神经元, 运动障碍, 神经祖细胞, 转录因子

Abstract: The generation of human induced pluripotent stem cell–derived motor neurons overcomes limited access to human tissues and offers an unprecedented approach to modeling motor neuron diseases such as dystonia and amyotrophic lateral sclerosis. Motor neurons generated through different strategies may exhibit substantial differences in purity, maturation, characterization, and even neuronal identity, leading to variable outcomes in disease modeling and drug screening. However, very few comparative studies have been conducted to determine the similarities and differences among motor neurons prepared via different protocols. In this study, we prepared human induced pluripotent stem cell–derived motor neurons via lentiviral delivery of transcription factors and chemical induction and performed a systematic comparative analysis. We found that motor neurons generated by both approaches showed typical motor neuron morphology and robustly expressed motor neuron-specific markers, such as nuclear homeobox transcription factor 9 and choline acetyltransferase. The chemical induction protocol utilizes a combination of small molecules to induce motor neuron differentiation, offering a significantly faster maturation time of 35 days compared to 46 days with lentiviral delivery of transcription factors. Additionally, while lentiviral delivery of transcription factors are suitable for downstream biochemical analysis, chemical induction are more applicable for therapeutic approaches as they avoid the use of lentiviruses. Both approaches produce motor neurons with high purity (> 95%) and yield. No significant differences were found between chemical induction and lentiviral delivery of transcription factors in terms of motor neuron markers and maturation markers. These robust methodologies offer researchers powerful tools for investigating motor neuron diseases and potential therapeutic strategies.

Key words: chemicals, human induced pluripotent stem cells, lentivirus, motor neuron diseases, motor neurons, movement disorders, neural progenitor cells, transcription factors